title: Determination of time-dependent accumulation of D-lactate in the streptozotocin-induced diabetic rat kidney by column-switching HPLC with fluorescence detection.
abstract: For better understanding the complete metabolism and the physiological role of D-lactate, the concentrations of D-lactate in the serum, liver and kidney of normal and diabetic rats were determined by our established column-switching HPLC method with pre-column fluorescence derivatization. Eight-week-old male Sprague-Dawley rats were administered with streptozotocin (STZ) (80 mg/kg) or citrate buffer intraperitoneally. The tissues were then removed and homogenized after 4, 8, 12 and 16 weeks of drug administration, respectively. The homogenates were centrifuged at 1200 × g for 10 min, then the supernatants were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ), separated on an ODS column followed by a Chiralpak AD-RH chiral column for enantioseparation. The results showed that the D-lactate content elevated in all the 3 examined tissues under diabetic stages. In addition, D-lactate concentrations in rat kidney were accumulated significantly and time-dependently in diabetic groups after receiving STZ for 4, 8, 12 and 16 weeks (2.99, 13.11, 18.19, 23.23 vs. 0.79 μmol/mg protein as control group). Moreover, the kidney of induced 12-week diabetic rat renal showed some histological changes of progressive diabetic nephropathy. The results suggest that D-lactate may be used as a marker of diabetic nephropathy.
authors.affiliation: School of Pharmacy, College of Pharmacy, Taipei Medical University, 250 Wuxing St., Taipei 11031, Taiwan.
authors.forename: Mei-Hsiang
authors.initials: MH
authors.lastname: Lin
authors.forename: Hsiang-Yin
authors.initials: HY
authors.lastname: Chen
authors.forename: Tzu-Hsin
authors.initials: TH
authors.lastname: Liao
authors.forename: Tzu-Chuan
authors.initials: TC
authors.lastname: Huang
authors.forename: Chien-Ming
authors.initials: CM
authors.lastname: Chen
authors.forename: Jen-Ai
authors.initials: JA
authors.lastname: Lee
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D006651:Histocytochemistry; D007668:Kidney / Q000737:chemistry* / Q000378:metabolism; D007672:Kidney Cortex / Q000473:pathology; D019344:Lactic Acid / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D016014:Linear Models; D008297:Male; D011765:Pyruvaldehyde / Q000032:analysis / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism; D013311:Streptozocin; D013997:Time Factors
chemical_list: D015415:Biomarkers; D019344:Lactic Acid; D013311:Streptozocin; D011765:Pyruvaldehyde